Investigation on the properties and kinetics of glucose-fed aerobic granular sludge

Aerobic granulation is a process in which suspended biomass aggregate and form discrete well-defined granules in aerobic systems. To investigate the properties and kinetics of aerobic granular sludge, aerobic granules were cultivated with glucose synthetic wastewater in a series of sequencing batch reactors (SBR). The spherical shaped granules were observed on 8th day with the mean diameter of 0.1 mm. With the organic loading rate (OLR) being increased to 4.0 g COD L⁻¹ d⁻¹, aerobic granules grew matured with spherical shape. The size of granules ranged from 1.2 to 1.8 mm, and the corresponding settling velocity of individual granule was 24.2–36.4 m h⁻¹. The oxygen utilization rate (OUR) of mature granules was 41.90 g O₂ kg MLSS⁻¹ h⁻¹, which was two times higher than that of activated sludge (18.32 g O₂ kg MLSS⁻¹ h⁻¹). The experimental data indicated that the substrate utilization and biomass growth kinetics generally followed Monod's kinetics model. The corresponding kinetic coefficients of k (maximum specific substrate utilization rate), K_s (half velocity coefficient), Y (growth yield coefficient) and K_d (decay coefficient) were determined as follows, k_c = 23.65 d⁻¹, K_c = 3367.05 mg L⁻¹, K_N = 0.038 d⁻¹, K_N = 29.65 mg L⁻¹, Y = 0.1927–0.2022 mg MMLS (mg COD)⁻¹ and K_d = 0.00845–0.0135 d⁻¹, respectively. Those properties of aerobic granules made aerobic granules system had a short setup period, high substrate utilization rate and low sludge production.     

The investigation of dairy industry wastewater treatment in a biological high performance membrane system

The dairy industry is generally considered to be the largest source of food processing wastewater in many countries. The highly variable nature of dairy wastewaters in terms of volumes and flowrates and in terms of high organic materials contents such as COD 921–9004 mg L⁻¹, BOD 483–6080 mg L⁻¹, TN of 8–230 mg L⁻¹ and SS of 134–804 mg L⁻¹ makes the choice of an effective wastewater treatment regime difficult. A high performance bioreactor, an aerobic jet loop reactor, combined with a ceramic membrane filtration unit, was used to investigate its suitability for the treatment of the dairy processing wastewater. The oxygen transfer rates of the bioreactor were found to be very high (100–285 h⁻¹) on the operating conditions. A loading rate of 53 kg COD m⁻³ d⁻¹ resulted in 97–98% COD removal efficiencies under 3 h hydraulic retention time. The high MLSS concentrations could be retained in the system (up to 38,000 mg L⁻¹) with the contribution of UF (ultrafiltration) unit. During the filtration of activated sludge, the fluxes decreased with increasing MLSS. Cake formation fouling was determined as dominant fouling mechanisms. The results demonstrate that jet loop membrane bioreactor system was a suitable and effective treatment choice for treating dairy industry wastewater.     

Development and examination of a granular nitrogen-fixing wastewater treatment system

This work presents the first success at aerobic granulation in a nitrogen deficient system. Two sequencing batch reactors (SBRs) were used to treat nitrogen deficient (the N-fix system) or nitrogen-sufficient (containing NH₄Cl) synthetic wastewater (acetic acid as the sole carbon source). Granulation was observed in both systems, with particularly large granules (average diameter: 7 mm) grown in the N-fix system. We propose that the unique morphology of nitrogen-fixing granules is a consequence of the response of oxygen-sensitive diazotrophs to elevated oxygen concentrations.Both the nitrogen-fixing and nitrogen-supplemented systems were shown to be capable of removing all of the influent substrate carbon. Excellent biomass settleability characteristics were obtained, with the N-fix system having a final sludge volume index (SVI) of less than 100 mL g⁻¹ and its granules having settling velocities of over 1.4 cm s⁻¹. However, moderately high solids discharges were recorded for both systems, which revealed a potential limitation of granular sludge processes that is not widely discussed in the literature.     

Aerobic granulation in sequencing batch reactors with different reactor height/diameter ratios

Effects of reactor height/diameter ratios ranged from 24 to 4 corresponding to reactor settling velocities from 12 to 2 m h^?1 on aerobic granulation were investigated. It was found that granules appeared after 1-week operation and granule volume percentages exceeded 50% after 2–3 weeks in four reactors. In addition, similar granule fraction of 94–96% was found at steady state in all four reactors. Sludge volume index (SVI), average sludge size, biomass density and granule settling velocity at steady state were around 50 ml g^?1, 1800 μm, 53 g l^?1 and 40 m h^?1, respectively, in four reactors. Extracellular polymeric substances (EPS) and specific oxygen uptake rate (SOUR) were around 38 mg g^?1 VSS and 40 mg O₂ g^?1 VSS h^?1, respectively. Denaturing gradient gel electrophoresis (DGGE) fingerprint of sludge in four reactors showed the same microbial population shift during the start-up period and same microbial community structure during steady-state period. These results recommended strongly that reactor height/diameter ratio or reactor setting velocity in the used range in this study did not affect granule formation, physical characteristics, microbial community structure of granules and stable operation of granular sludge reactor. Reactor height/diameter ratio thus can be very flexible in the practice, which is important for the application of aerobic granule technology.     

Biological treatment of low-salinity shrimp aquaculture wastewater using sequencing batch reactor

In order to improve the water quality in shrimp aquaculture operated under low-salinity conditions, a sequencing batch reactor (SBR) was tested for treatment of the wastewater. This water from the backwash of a single-bead filter from the Waddell Mariculture Center, South Carolina, contained high concentrations of carbon and nitrogen and was successfully treated using the SBR. By operating the reactor sequentially in aerobic, anoxic and aerobic modes, nitrification and denitrification were achieved, as well as removal of carbon. Specifically, the initial chemical oxygen demand (COD) concentration of 1201 mg l⁻¹ was reduced to 32 mg l⁻¹ within 8 days of reactor operation. Ammonia in the sludge was nitrified within 3 days. The denitrification of nitrate was achieved by the anoxic process and total removal of nitrate was observed.     

Dominance of yeast in activated sludge under acidic pH and high organic loading

Flow cytometry-fluorescent in situ hybridization (FC-FISH) was used to investigate the effect of controlled pH and/or varied organic loading on the content of yeast and bacterial cells in an activated sludge system (AS) individually operating in continuous and batch mode for treatment of high-strength industrial wastewater. Specifically, we attempted to develop a yeast-predominant activated sludge system (Y-AS). For the batch-mode AS, bacteria-dominated AS (B-AS) obtained at pH 6.5–7.5 induced higher chemical oxygen demand (COD) removal than Y-AS obtained at acidic pH (5.0–6.0 and 4.0–5.0). For the continuous-mode AS operating at COD loadings of 2.5–2.8 kg COD m⁻³ d⁻¹, it was difficult to achieve a Y-AS solely by controlling the pH level at 7.0 to 5.1 then to 4.1 because bacteria stably accounted for greater than 98% of the total cells, regardless of the pH levels. Therefore, the effects of varied COD loadings (2.1, 8.7 and 21.0 kg COD m⁻³ d⁻¹) on continuous-mode AS operation at acidic pH (4.5) was investigated. Both acidic pH and high COD loading levels were found to be prerequisites for yeast to dominate the sludge microbial community in the continuous-mode AS.     

Dynamic simulation of a WWTP operated at low dissolved oxygen condition by integrating activated sludge model and a floc model

While an aeration tank in an activated sludge process is often operated with high dissolved oxygen (DO) concentration to ensure organic degradation and nitrification, it may be operated at low DO concentration to reduce energy consumption and achieve desired denitrification. The ASM1 (Activated Sludge Model No. 1) can be used to describe the activated sludge process if the nitrification and denitrification occur either during different phases or in different tanks, but it may encounter problems in simulating the denitrification phenomenon caused by low DO concentration in the aeration tank. In the present work, we developed a model integrating the ASM1 kinetics and a biofloc model to account for the actual anoxic and aerobic rates. Oxygen was assumed the only substrate of both bio-kinetically and flux limiting in the flocs and its dispersion coefficient was estimated as 1.2 × 10⁻⁴ m² day⁻¹ by using a set of measured effluent qualities of a full-scale wastewater treatment plant (WWTP) operating at low DO concentration (∼0.80 mg L⁻¹) for 60 days. Simulation studies predicted the optimal DO level of 0.36 mg L⁻¹ which would lead to minimum total nitrogen of 15.7 mg N L⁻¹ and also showed the insignificance of the addition of carbon source for nitrogen removal for the operation under study. The developed model may be helpful for process engineers to predict the plant behaviors under various configurations or operating strategies.     

A key cultivation technology for denitrifying granular sludge

Well-formed denitrifying granular sludge with a biomass concentration of 24.8 gVSS L^?1 and a specific nitrate removal rate of 0.19 gNO₃-N gVSS^?1 d^?1 was obtained in an upflow sludge blanket (USB) reactor by cultivating seeded aerobic flocculent sludge for 6–8 weeks. Regularity phenomena exist in the granulation including flotation of flocculent sludge, formation of fine granules, occurrence of channelling, and formation of mature granular sludge. The granulation is similar to crystal growth, that the non-denitrifying bacteria evolve into the carriers (fine granules), on the surface of which denitrifying bacteria proliferate and develop into mature granular sludge.There are several key parameters that must be considered when developing a good denitrifying granular sludge. First, the proper seed sludge must be chosen (VSS/SS at 0.65–0.75, SRT over 25 days) to accelerate the granulation process. Secondly, any floating sludge should be stirred, and the sludge loading rate should be within the range of 0.05–0.15 gNO₃-N gVSS^?1 d^?1 until fine granules emerge. Additionally, spontaneous gas agitation or interval air-blowing should be used to effectively eliminate channelling; Finally, the sludge loading rate should be less than 0.25 gNO₃-N gVSS^?1 d^?1 until dense, mature granular sludge appears. This study could support and promote the full-scale application of denitrifying granular sludge.     

A phenanthrene-degrading strain Sphingomonas sp. GY2B isolated from contaminated soils

This study systematically characterized an aerobic bacterial strain Sphingomonas sp. GY2B for biotransformation of phenanthrene. The strain was isolated from soils contaminated with polycyclic aromatic hydrocarbons (PAHs) and was shown to efficiently use phenanthrene as the sole carbon and energy source. The antibiotics discs susceptibility test revealed that the bacterium was susceptible to some commonly used antibiotics, such as cefuroxime, chloramphenicol, erythromycin and tetracycline. It showed better growth at pH 7.4 and 30 °C and in a mineral salts medium (MSM) with phenanthrene at 100 mg L⁻¹ as the substrate. The results indicated that 99.8% of the substrate had been degraded and that salicylate route was likely the metabolic pathway. When added as the second organic chemical, glucose could enhance the bacterial growth at low concentration (10–200 mg L⁻¹), but could inhibit cell growth at high concentration (>500 mg L⁻¹). Further study showed that strain GY2B could also use naphthalene, phenol, 1-hydroxy-2-naphthoic acid, 2-naphthol, salicylic acid and catechol as the sole carbon and energy source, but did not grow on 1-naphthol which could be co-metabolized in the present of phenanthrene or 1-hydroxy-2-naphthoic acid.     

Purification and characterization of cyanogenic β-glucosidase from wild apricot (Prunus armeniaca L.)

β-Glucosidase catalyzes the sequential breakdown of cyanogenic glycosides in cyanogenic plants. The β-glucosidase from Prunus armeniaca L. was purified to 8-fold, and 20% yield was obtained, with a specific activity of 281 U/mg protein. The enzyme showed maximum activity in 0.15 M sodium citrate buffer, pH 6, at 35 °C with p-nitrophenylglucopyranoside as substrate. The β-glucosidase from wild apricot was used successfully for the saccharification of cellobiose into D-glucose. This enzyme has a V_max of 131.6 μmol min⁻¹ mg⁻¹ protein, K_m of 0.158 mM, K_cat of 144.8 s⁻¹, K_cat/K_m of 917.4 mM⁻¹ s⁻¹, and K_m/V_max of 0.0012 mM min mg μmole⁻¹, using cellobiose as substrate. The half-life, deactivation rate coefficient, and activation energy of this β-glucosidase were 12.76 h, 1.509 × 10⁻⁵ s⁻¹, and 37.55 kJ/mol, respectively. These results showed that P. armeniaca is a potential source of β-glucosidase, with high affinity and catalytic capability for the saccharification of cellulosic material.     

Biodegradation kinetics of stored wastewater substrates by a mixed microbial culture

The anaerobic accumulation of several organic pollutants from industrial wastewaters, as storage substrates, and their subsequent aerobic biodegradation using a wastewater treatment mixed microbial culture for biological nutrient removal has been studied. The amount and the kinetics of substrate accumulation in the anaerobic stage depended on the characteristics of the wastewater fed to the anaerobic stage. Depending on the substrate used, levels of between 27 and 86% of storage polymers were accumulated with respect to the level obtained on feeding with acetate. The biodegradation kinetics were studied by modelling respirometry results. During the aerobic stage, oxygen-consumption data obtained in the respirometric tests were fitted to a model using a non-linear fitting estimation method. The simulation data obtained correlated well with the experimental oxygen-consumption data. The estimated kinetic parameters obtained indicate that each storage polymer was degraded at a different rate. However, the values obtained for the storage polymer half-saturation coefficient, K_S: 16 mg COD l⁻¹, and for the coefficient for endogenous respiration, b: 0.008 h⁻¹, were similar in all the experiments. The results indicate that each substrate produces the synthesis of a specific storage polymer that is degraded at a different rate.     

Urine nitrification with a synthetic microbial community

During long-term extra-terrestrial missions, food is limited and waste is generated. By recycling valuable nutrients from this waste via regenerative life support systems, food can be produced in space. Astronauts’ urine can, for instance, be nitrified by micro-organisms into a liquid nitrate fertilizer for plant growth in space. Due to stringent conditions in space, microbial communities need to be be defined (gnotobiotic); therefore, synthetic rather than mixed microbial communities are preferred. For urine nitrification, synthetic communities face challenges, such as from salinity, ureolysis, and organics.In this study, a synthetic microbial community containing an AOB (Nitrosomonas europaea), NOB (Nitrobacter winogradskyi), and three ureolytic heterotrophs (Pseudomonas fluorescens, Acidovorax delafieldii, and Delftia acidovorans) was compiled and evaluated for these challenges. In reactor 1, salt adaptation of the ammonium-fed AOB and NOB co-culture was possible up to 45 mS cm⁻¹, which resembled undiluted nitrified urine, while maintaining a 44 ± 10 mg NH₄⁺–N L⁻¹ d⁻¹ removal rate. In reactor 2, the nitrifiers and ureolytic heterotrophs were fed with urine and achieved a 15 ± 6 mg NO₃⁻–N L⁻¹ d⁻¹ production rate for 1% and 10% synthetic and fresh real urine, respectively. Batch activity tests with this community using fresh real urine even reached 29 ± 3 mg N L⁻¹ d⁻¹. Organics removal in the reactor (69 ± 15%) should be optimized to generate a nitrate fertilizer for future space applications.     

Operation of partial nitrification to nitrite of landfill leachate and its performance with respect to different oxygen conditions

The coupled system of partial nitrification and anaerobic ammonium oxidation (Anammox) is efficient in nitrogen removal from wastewater. In this study, the effect of different oxygen concentrations on partial nitrification performance with a sequencing batch reactor (SBR) was investigated. Results indicate that, partial nitrification of landfill leachate could be successfully achieved under the 1.0–2.0 mg L⁻¹ dissolved oxygen (DO) condition after 118 d long-term operation, and that the effluent is suitable for an Anammox reactor. Further decreasing or increasing the DO concentration, however, would lead to a decay of nitrification performance. Additionally, the MLSS concentration in the reactor increased with increasing DO concentration. Respirometric assays suggest that low DO conditions (<2 mg L⁻¹) favor the ammonia-oxidizing bacteria (AOB) and significantly inhibit nitrite oxidizing bacteria (NOB) and aerobic heterotrophic bacteria (AHB); whereas high DO conditions (>3 mg L⁻¹) allow AHB to dominate and significantly inhibit AOB. Therefore, the optimal condition for partial nitrification of landfill leachate is 1.0–2.0 mg L⁻¹ DO concentration.     

Purification and characterization of an exo-polygalacturonase produced by Penicillium viridicatum RFC3 in solid-state fermentation

The pectinolytic enzyme obtained from Penicillium viridicatum RFC by solid-state fermentation was purified to homogeneity by pretreatment with kaolin (40 mg mL⁻¹) and ultrafiltration, followed by chromatography on a Sephadex G50 column. The apparent molecular weight of the enzyme was 24 kDa. Maximal activity occurred at pH 6.0 and at 60 °C. The enzyme proved to be an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of highly esterified pectin. The presence of 10 mM Ba²⁺ increased the enzyme activity by 96% and its thermal stability by 30%, besides increasing its stability at acid pH. The apparent K_m with apple pectin as substrate was 1.82 mg mL⁻¹ and the V_max was 81 μmol min⁻¹ mg⁻¹.     

Naphthalene and biphenyl oxidation by two marine Pseudomonas strains isolated from Venice Lagoon sediment

A sediment sample from Venice Lagoon was found to be contaminated with 475 mg Kg⁻¹ polycyclic aromatic hydrocarbons (PAHs). Naphthalene was the principal pollutant at 26% of total PAHs. Two strains of Pseudomonas SN1 and SB1 were isolated from sediment amended with 2% naphthalene. 16S rRNA gene sequence analysis indicated that the two strains have about 99% nucleotide identity with strains of the genus Pseudomonas, and are very close to Pseudomonas stutzeri. Their metabolic profiles showed significant nutritional differences, the most significant of which was that SN1 grows in marine mineral medium spiked with naphthalene and SB1 grows with biphenyl as sole carbon and energy sources. Pseudomonas sp. SN1 had a doubling time of 3.1 h with 2% naphthalene and SB1 had a doubling time of 19.5 h with 2% biphenyl. Strain SN1 oxidised naphthalene at 564±32 mg O₂ l⁻¹ d⁻¹ and SB1 oxidised biphenyl at 426±25 mg O₂ l⁻¹ d⁻¹ in respirometry reaction vessels under controlled conditions. Screening of the two strains for dioxygenase genes involved in the first step of the two hydrocarbon degradation pathways, by polymerase chain reaction, showed naphthalene dioxygenase in SN1 and biphenyl dioxygenase in SB1. The strains each have a different catechol 2,3-dioxygenase responsible for cleavage of the aromatic ring.     

Protocorm culture of Dendrobium candidum in balloon type bubble bioreactors

Protocorm cultures of Dendrobium candidum were established in balloon type bubble bioreactors using Murashige and Skoog (MS) medium with 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA), 2.5% (w/v) sucrose, 5:25 mM NH₄:NO₃ and 1% (v/v) banana homogenate for the production of biomass and bioactive compounds. In 3 l bioreactor containing 2 l medium, a maximum protocorm biomass (21.0 g l⁻¹ dry biomass) and also optimum quantities of total polysaccharides (389.3 mg g⁻¹ DW), coumarins (18.0 mg g⁻¹ DW), polyphenolics (11.9 mg g⁻¹ DW), and flavonoids (4.5 mg g⁻¹ DW) were achieved after 7 weeks of culture. Based on these studies, 5 and 10 l bioreactor cultures were established to harvest 80 g and 160 g dry biomass. In 10 l bioreactors, the protocorms grown were accumulated with optimal levels of polysaccharides (424.1 mg g⁻¹ DW), coumarins (15.8 mg g⁻¹ DW), polyphenols (9.03 mg g⁻¹ DW) and flavonoids (4.7 mg g⁻¹ DW). The bioreactor technology developed here will be useful for the production of important bioactive compounds from D. candidum.     

Inhibition kinetics of a commercial mixed culture by ammonium sulfate

The inhibitory effect of ammonium sulfate on a commercial mixed culture, used in biological waste-water treatment was studied under aerobic batch conditions. Several mathematical models of enzyme and growth kinetics including a death factor were analyzed through nonlinear regression to find the best fit to corresponding data of inhibition. The best fit model was found to be the generalized Monod type with a death factor having the biokinetic parameters; μ_max 0.681 h⁻¹, K_s 0.224 g dm⁻³, K_i 56240 g dm⁻³, K 0.055 g dm⁻³ and k_d 0.052 h⁻¹ to represent the experimental data accurately. The low saturation coefficient value along with high maximum specific growth rate and inhibition coefficient denotes the competitive characteristics of commercial mixed cultures in the biological treatment of high ammonium polluted waste waters.     

Comparative kinetic study between moving bed biofilm reactor-membrane bioreactor and membrane bioreactor systems and their influence on organic matter and nutrients removal

New technologies regarding wastewater treatment have been developed. Among these technologies, the moving bed biofilm reactor combined with membrane bioreactor (MBBR-MBR) is a recent solution alternative to conventional processes. This paper presents the results obtained from three wastewater treatment plants working in parallel. The first wastewater treatment plant consisted of a membrane bioreactor (MBR), the second one was a MBBR-MBR system containing carriers both in anoxic and aerobic zones, and the last one consisted of a MBBR-MBR system which contained carriers only in the aerobic zone. The reactors operated with a hydraulic retention time of 26.47 h. During the study, the difference between the experimental plants was not statistically significant concerning organic matter and nutrients removal. However, different tendencies regarding nutrients removal are shown by the three wastewater treatment plants. In this sense, the performances in terms of nitrogen and phosphorus removal of the MBBR-MBR system which contained carriers only in the aerobic zone (67.34 ± 11.22% and 50.65 ± 11.13%, respectively) were slightly better than those obtained from another experimental plants. As a whole, the pilot plant which consisted of a MBR showed better performance from the point of view of the kinetics of the heterotrophic and autotrophic biomass with values of μ_m,H = 0.00858 h⁻¹, μ_m,A = 0.07646 h⁻¹, K_M = 2.37 mg O₂ L⁻¹ and K_NH = 1.31 mg N L⁻¹.     

Chromate reduction by resting cells of Achromobacter sp. Ch-1 under aerobic conditions

Chromate reduction was carried out by resting cells of Achromobacter sp. Ch-1 with lactate as electron donor under aerobic conditions. The reduction activity of the samples supplemented with lactate was two times as those without lactate. The reduction rate was influenced by initial pH and lactate concentration. Under the optimal conditions, pH 9.0 and 4000 mg l⁻¹ lactate supplement, reduction rate was 5.45 mg l⁻¹ min⁻¹. The reduction rate decreased with increasing of Cr(VI) concentrations and increased with cell densities proportionally. The maximum reduction limit of Ch-1 cells was obtained at 2107 mg l⁻¹ of Cr(VI).     

Cultivation of Chlorella vulgaris in tubular photobioreactors: A lipid source for biodiesel production

Chlorella vulgaris was cultivated in two different 2.0 L-helicoidal and horizontal photobioreactors at 5 klux using the bicarbonate contained in the medium and ambient air as the main CO₂ sources. The influence of bicarbonate concentration on biomass growth as well as lipid content and profile was first investigated in shake flasks, where the stationary phase was achieved in about one half the time required by the control. The best NaHCO₃ concentration (0.2 g L⁻¹) was then used in both photobioreactors. While the fed-batch run performed in the helicoidal photobioreactor provided the best result in terms of biomass productivity, which was (84.8 mg L⁻¹ d⁻¹) about 2.5-fold that of the batch run, the horizontal configuration ensured the highest lipid productivity (10.3 mg L⁻¹ d⁻¹) because of a higher lipid content of biomass (22.8%). These preliminary results suggest that the photobioreactor configuration is a key factor either for the growth or the composition of this microalga. The lipid quality of C. vulgaris biomass grown in both photobioreactors is expected to meet the standards for biodiesel, especially in the case of the helicoidal configuration, provided that further efforts will be made to optimize the conditions for its production as a biodiesel source.